Abstract
The aberrant expression of mRNAs participates in the pathogenesis of hepatic fibrosis. However, the precise mechanisms regulated by microRNAs (miRNAs) remain unclear. This study aims to investigate the functions about differentially expressed mRNAs (DEMs) in liver fibrosis and their regulatory mechanisms. The DEMs datasets about hepatic stellate cells (HSCs) obtained from hepatic fibrosis mice versus HSCs obtained from normal mice were downloaded from the GEO database (GSE120281). According to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of the GSE120281 datasets, ECM-receptor interaction was the most significant enrichment pathway that was correlated with hepatic fibrosis, and the fibronectin 1 (FN1) gene was upregulated most significantly in the signaling pathway. Downregulation of the expression of the FN1 gene by transfecting with FN1-siRNA alleviated the activity of HSCs. Four different bioinformatics web-based tools were used to predict that microRNA-96-5p (miR-96-5p) would directly target FN1, and a luciferase assay further confirmed this. Moreover, miR-96-5p was declined in activated HSCs and FN1, whereas laminin γ1 (LAMC1), collagen 1α1 (COL1A1) in the ECM-receptor interaction pathway, and the fibrosis marker α-smooth muscle actin (α-SMA) could be reduced by upregulation of the miRNA. Additionally, miR-96-5p expression was low in CCl4-induced liver fibrosis mice. Increased miR-96-5p expression alleviated liver fibrosis, improved liver function, and inhibited the expression of α-SMA, FN1, COL1A1, and LAMC1. In conclusion, this study indicated that upregulation of miR-96-5p could reduce HSC activation and relieve hepatic fibrosis by restraining the FN1/ECM-receptor interaction pathway.
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All data and materials used during this study are available from the corresponding author.
Abbreviations
- ALT:
-
alanine aminotransferase
- AST:
-
aspartate aminotransferase
- α-SMA:
-
α-smooth muscle actin
- COL1A1:
-
collagen 1α1
- DEMs:
-
differentially expressed mRNAs
- ECM:
-
extracellular matrix
- FN1:
-
fibronectin 1
- GO:
-
geneontology
- HSC:
-
hepatic stellate cell
- IHC:
-
immunohistochemistry
- KEGG:
-
kyoto encyclopedia of genes and genomes
- LAMC1:
-
laminin γ1
- miRNA:
-
microRNA
- MUT:
-
mutant
- NC:
-
negative control
- qRT-PCR:
-
quantitative real-time PCR
- TGF-β1:
-
transforming growth factor-β1
- WT:
-
wild-type
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The study was funded by the Social Development Fund of Zhenjiang (No. SH2020027).
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YZ and TG performed the experiment and drafted the manuscript. SX and JW analyzed and interpreted the data. XZ and JW designed the experiment. SX funded the experiment. XZ reviewed and edited the manuscript.
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Protocols for animal experiments were approved by the Institutional Animal Care and Use Committee of Jiangsu University (protocol code: UJS-IACUC-2021030269).
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Zhang, Y., Gu, T., Xu, S. et al. Anti-Liver Fibrosis Role of miRNA-96-5p via Targeting FN1 and Inhibiting ECM-Receptor Interaction Pathway. Appl Biochem Biotechnol 195, 6840–6855 (2023). https://doi.org/10.1007/s12010-023-04385-1
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DOI: https://doi.org/10.1007/s12010-023-04385-1